11,391 results match your criteria Journal of Immunological Methods [Journal]


Biological variation of peripheral blood T-lymphocytes.

J Immunol Methods 2019 Apr 22. Epub 2019 Apr 22.

Ankara Education and Trainning Hospital, Biochemistry Department, Ankara, Turkey.

Background: Flow cytometric analysis of the lymphocyte subsets has become one of the most commonly used techniques in the routine clinical laboratory. It is frequently used in monitoring lymphocyte recovery after hematopoietic stem cell transplantation (HSCT), as well as diagnosis and treatment of acquired immunodeficiency syndrome (AIDS). Reliable biological variation (BV) data is needed for safe clinical application of these tests. Read More

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http://dx.doi.org/10.1016/j.jim.2019.04.002DOI Listing

Development of a novel multiplex electrochemiluminescent-based immunoassay to aid enterotoxigenic Escherichia coli vaccine development and evaluations.

J Immunol Methods 2019 Apr 17. Epub 2019 Apr 17.

Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA.

Enterotoxigenic Escherichia coli (ETEC) is a leading cause of bacterial diarrhea both among children in low and middle income countries and in travelers to these regions. Although there are several approaches to develop an effective vaccine for ETEC, no licensed vaccines are currently available. The most advanced ETEC vaccine candidates include multiple colonization factors along with the heat labile toxin B subunit. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183040
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http://dx.doi.org/10.1016/j.jim.2019.04.003DOI Listing
April 2019
1 Read

Development of a novel immunoassay for the simple and fast quantitation of neutrophil gelatinase-associated lipocalin using europium(III) chelate microparticles and magnetic beads.

J Immunol Methods 2019 Apr 17. Epub 2019 Apr 17.

Institute of Antibody Engineering, School of Laboratory Medicine and Biotechnology, Southern Medical University, 510515 Guangzhou, China. Electronic address:

Neutrophil gelatinase-associated lipocalin (NGAL) is a promising biomarker for diagnosing acute kidney injury (AKI). Currently, there are few assays for determining NGAL and they are complex, time-consuming or expensive. We aimed to establish an efficient immunoassay to measure NGAL in human urine simply and rapidly. Read More

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http://dx.doi.org/10.1016/j.jim.2019.04.004DOI Listing
April 2019
1 Read

Visualization and quantification of NK cell-mediated cytotoxicity over extended time periods by image cytometry.

J Immunol Methods 2019 Apr 2. Epub 2019 Apr 2.

Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA 02115, United States of America.

Natural killer (NK) cell-mediated cytotoxicity is traditionally measured using the chromium release assay, which measures the fraction of radioactive Cr released from dying target cells co-cultured with NK cells. However, the time frame of Cr release assays is limited to approximately 4 h due to spontaneous release of Cr. In the tumor microenvironment, interactions between NK cells and tumor cells occur over extended time periods, and NK cell-mediated cytotoxicity is modulated by cytokines produced by tumor cells and other immune cells. Read More

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http://dx.doi.org/10.1016/j.jim.2019.04.001DOI Listing

Production of monoclonal antibodies for measuring Avastin and its biosimilar by Sandwich ELISA.

J Immunol Methods 2019 Mar 31. Epub 2019 Mar 31.

AbMax Biotechnology Co., LTD, Beijing, China. Electronic address:

The development of Bevacizumab (Avastin) biosimilar products has grown rapidly over the last ten years as the original Avastin's patent will expire soon. The approval of Avastin biosimilars requires the demonstration of similarity between the biosimilar and the reference product. To support pre-clinical and clinical studies, pharmacokinetic (PK) assays are required to measure the biosimilar and Avastin with comparable precision and accuracy. Read More

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http://dx.doi.org/10.1016/j.jim.2019.03.013DOI Listing
March 2019
2 Reads

ELISA methods comparison for the detection of auto-antibodies against apolipoprotein A1.

J Immunol Methods 2019 Mar 27. Epub 2019 Mar 27.

Division of Laboratory Medicine, Diagnostic Department, Geneva University Hospitals, 4 rue Gabrielle-Perret-Gentil, 1205 Geneva, Switzerland; Department of Medical Specialties, Faculty of Medicine, University of Geneva, 1 rue Michel Servet, 1205 Geneva, Switzerland.

Background: Autoantibodies against apolipoprotein A1 (anti-apoA1 IgG) have emerged as an independent biomarker for cardiovascular disease and mortality. Across studies, different ELISA methods have been used to measure the level of circulating anti-apoA1 IgG which could lead to substantial result differences between assays.

Objectives: To make a comparative study of available anti-apoA1 IgG detection methods and to determine whether the choice of matrix sample (serum vs plasma) could influence the results. Read More

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http://dx.doi.org/10.1016/j.jim.2019.03.011DOI Listing

Eight-color panel for immune phenotype monitoring by flow cytometry.

J Immunol Methods 2019 May 23;468:40-48. Epub 2019 Mar 23.

Division of Clinical Analysis, Flow Cytometry Service, University Hospital of the Federal University of Santa Catarina (UFSC), Florianopolis, SC 88040-900, Brazil; Postgraduate Program in Pharmacy of the Federal University of Santa Catarina (UFSC), Florianopolis, SC 88040-900, Brazil; Clinical Analysis Department, Health Sciences Center, Federal University of Santa Catarina (UFSC), Florianopolis, SC 88040-900, Brazil. Electronic address:

Flow cytometry (FC) is a fast and highly informative technology that has gained prominence in immune phenotype monitoring. FC standardization is crucial to obtain reliable results that are comparable among laboratories and immune monitoring studies, as this method is influenced by several variables, including equipment, reagents, staining procedures, and pre-analytical and analytical factors. Recent studies have standardized antibody panels and analytical procedures to analyze circulating immune cells in peripheral blood (PB). Read More

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http://dx.doi.org/10.1016/j.jim.2019.03.010DOI Listing
May 2019
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The use of BIOCHIP technique in diagnosis of different types of pemphigus: Vulgaris and foliaceus.

J Immunol Methods 2019 May 22;468:35-39. Epub 2019 Mar 22.

Department of Dermatology and Immunodermatology, Medical University of Warsaw, Poland.

Background: Pemphigus is a rare, autoimmune blistering disease characterized by autoantibodies against desmoglein 3 (Dsg3) and 1 (Dsg1) with mucosal and/or skin involvement. Main types of pemphigus include mucosal pemphigus vulgaris (m-PV), mucocutaneus pemphigus vulgaris (mc-PV) and pemphigus foliaceus (PF) determined by clinical picture, positive direct and indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA).

Methods: We evaluated the sensitivity and specificity of a novel multi-substrate immunofluorescence technique called BIOCHIP in the diagnosis of main types of pemphigus. Read More

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http://dx.doi.org/10.1016/j.jim.2019.03.009DOI Listing

Development and characterization of a Zaire Ebola (ZEBOV) specific IgM ELISA.

J Immunol Methods 2019 May 22;468:29-34. Epub 2019 Mar 22.

Malaria Vaccine Branch, Walter Reed Army Institute of Research, Silver Spring, MD 20910, United States of America. Electronic address:

Immunoglobulin M (IgM) is the first antibody induced after the onset of an adaptive immune response against a pathogen or vaccine. Serological assays play a central role in evaluating these adaptive immunological responses. Such assays are not only crucial for the assessment of vaccine immunogenicity, but also inform on exposure to pathogens and cross-reactivity with other viruses. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759193004
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http://dx.doi.org/10.1016/j.jim.2019.03.008DOI Listing
May 2019
7 Reads
2.005 Impact Factor

Purification of IgG against ribonucleoprotein by a homemade immunoaffinity chromatography column for rabies diagnosis.

J Immunol Methods 2019 Mar 20. Epub 2019 Mar 20.

Instituto Pasteur, São Paulo, Brazil. Electronic address:

Polyclonal or monoclonal antibodies against rabies virus ribonucleoprotein (RNP) conjugated to fluorescein isothiocyanate (FITC) have been employed for Rabies virus (RABV) antigen detection by the direct fluorescent antibody test (DFA). To date, these biomolecules have been purified by traditional methods such as precipitation by ammonium sulfate or ion exchange chromatography followed by ammonium sulfate precipitation, which allows only for partial detection of the protein of interest. In this study, we aimed to purify anti-RNP polyclonal horse IgG antibodies by cation-exchange chromatography in combination with a homemade immunoaffinity chromatography on RNP immobilized (RNP-IAC). Read More

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http://dx.doi.org/10.1016/j.jim.2019.03.007DOI Listing
March 2019
2.005 Impact Factor

A standardised protocol for the evaluation of small extracellular vesicles in plasma by imaging flow cytometry.

J Immunol Methods 2019 May 16;468:61-66. Epub 2019 Mar 16.

School of Biomedical Sciences, University of Western Australia, Perth, WA, Australia; Stem Cell Unit, Institute for Respiratory Health, Perth, WA, Australia; Department of Respiratory Medicine, Fiona Stanley Hospital, Murdoch, WA, Australia.

Flow cytometry provides robust, multi-parametric and quantitative information on single cells which also exhibits enormous potential as a tool for small particle characterisation. Small extracellular vesicle (sEV) detection by flow cytometry remains compromised due to the high prevalence of swarm detection, which is defined by the simultaneous illumination of more than one sEV, recorded as a single event. Detection of sEVs by imaging flow cytometry presents a major advantage by having the ability to resolve single particles from swarm detection based on the image features recorded for each event. Read More

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http://dx.doi.org/10.1016/j.jim.2019.03.006DOI Listing

An IgY-based immunoassay to evaluate the biomarker potential of the Tannerella forsythia virulence factor karilysin in human saliva.

J Immunol Methods 2019 Mar 14. Epub 2019 Mar 14.

Department of Biochemistry and Molecular Biology, University of Southern Denmark, 5230 Odense M, Denmark; Research and Development Department, Sanovo Biotech A/S, DK-5000 Odense, Denmark.

Tannerella forsythia is a gram-negative anaerobic bacterium that is associated with the development of destructive periodontal disease. T. forsythia secretes the metalloprotease-like enzyme karilysin. Read More

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http://dx.doi.org/10.1016/j.jim.2019.03.003DOI Listing

Experimental protocol for development of adjuvant-free murine chronic model of allergic asthma.

J Immunol Methods 2019 May 14;468:10-19. Epub 2019 Mar 14.

National Research Center - Institute of immunology of Federal Medico-Biological Agency, 115478, 24, Kashirskoye Shosse, Moscow, Russia.

Background: Mouse models of allergic asthma play a crucial role in exploring of asthma pathogenesis and testing of novel anti-inflammatory drugs. Widely used acute asthma models usually developed with adjuvant (aluminum hydroxide (alum)) do not reproduce one of the main asthma feature - airway remodeling while chronic asthma model mimic the pathophysiology of human disease. Moreover, the use of alum causes distress in experimental animals and impedes the test of adjuvant-containing drugs. Read More

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http://dx.doi.org/10.1016/j.jim.2019.03.002DOI Listing
May 2019
1 Read

A novel method for determining antibody-dependent cellular phagocytosis.

J Immunol Methods 2019 May 14;468:55-60. Epub 2019 Mar 14.

Department of BioAnalytical Sciences, Genentech Inc, 1 DNA Way, South San Francisco, CA 94080-4990, United States. Electronic address:

Antibody-based therapeutics are powerful tools to treat disease. While their mechanism of action (MOA) always involves binding to a specific target via the Fab region of the antibody, the induction of effector functions through the Fc region of the antibody is equally important for antibody therapeutics designed to deplete tumor cells. By binding of the Fc region to Fc gamma receptors (FcγRs) on the surface of immune cells or complement factors, antibody therapeutics exert effector functions such as antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), both of which induce target cell death and aid in the efficacy of treatment. Read More

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http://dx.doi.org/10.1016/j.jim.2019.03.001DOI Listing
May 2019
2 Reads

Depletion of interfering IgG and IgM is critical to determine the role of IgE in pegvaliase-associated hypersensitivity.

J Immunol Methods 2019 May 14;468:20-28. Epub 2019 Mar 14.

BioMarin Pharmaceutical Inc., 105 Digital Drive, Novato, CA 94949, USA.

Pegvaliase is an enzyme substitution therapy developed to lower blood phenylalanine (Phe) in adults with phenylketonuria (PKU). In phase 3 clinical studies, pegvaliase substantially reduced mean blood Phe in adult subjects with PKU. The most common type of adverse event observed in the pegvaliase clinical program was hypersensitivity adverse events (HAEs), which included occurrences of arthralgia, rash, and pruritis. Read More

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http://dx.doi.org/10.1016/j.jim.2019.03.004DOI Listing
May 2019
4 Reads
2.005 Impact Factor

Performance evaluation of anti-fixed Leishmania infantum promastigotes immunoglobulin G (IgG) detected by flow cytometry as a diagnostic tool for visceral Leishmaniasis.

J Immunol Methods 2019 Feb 25. Epub 2019 Feb 25.

Fundação Oswaldo Cruz (Fiocruz-Pernambuco), Instituto Aggeu Magalhães, Recife, Pernambuco, Brazil.

Visceral Leishmaniasis (VL) is a severe disease, caused by the protozoans Leishmania infantum and L. donovani that is widely diagnosed using serological tools. These, however, have limitations in performance that limit their use for the correct identification of the cases. Read More

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http://dx.doi.org/10.1016/j.jim.2019.02.009DOI Listing
February 2019
1 Read

Identification of murine antigen-specific T follicular helper cells using an activation-induced marker assay.

J Immunol Methods 2019 Apr 22;467:48-57. Epub 2019 Feb 22.

Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, Victoria, Australia. Electronic address:

Protective antibody (Ab) responses induced by natural infection or vaccination play a central role in defense against invasive pathogens. Germinal centers (GCs) are the sites of Ab affinity maturation and T follicular helper (Tfh) cells are a critical factor for driving GC formation and B cell selection. Therefore characterization of antigen (Ag)-specific Tfh cells is increasingly essential to define the mechanistic basis of protective antibody responses. Read More

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http://dx.doi.org/10.1016/j.jim.2019.02.008DOI Listing
April 2019
2 Reads

Background autofluorescence induced by plant extracts in human lymphocytes: A flow cytometric analysis of a critical bias.

J Immunol Methods 2019 May 22;468:1-9. Epub 2019 Feb 22.

Laboratory of Immunology, Department of Pharmacy, Universidade Federal dos Vales do Jequitinhonha e Mucuri (UFVJM), Diamantina, Minas Gerais 39100-00, Brazil; Graduate Program in Pharmaceutical Sciences, Universidade Federal dos Vales do Jequitinhonha e Mucuri (UFVJM), Diamantina, Minas Gerais 39100-000, Brazil; Multicenter Graduate Program in Physiological Sciences/UFVJM, Universidade Federal dos Vales do Jequitinhonha e Mucuri (UFVJM), Diamantina, Minas Gerais 39100-000, Brazil. Electronic address:

The presence of background autofluorescence sources is considered as an important problem when performing fluorometric methods, due to the possible spectral overlap between it and the fluorescence emission of probes. Regarding that, we evaluated the presence of background autofluorescence in human lymphocytes after the treatment with extracts from three medicinal plants, including ethanolic extract from aerial parts of Ageratum fastigiatum, ethanolic extract from aerial parts of Eriosema campestre and the ethanolic extract from stem of Pseudobrickellia brasiliensis. Human peripheral blood mononuclear cells were treated with each extract in vitro during 24 h, followed by flow cytometric analysis. Read More

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http://dx.doi.org/10.1016/j.jim.2019.02.007DOI Listing

Development of a kinetic antibody-dependent cellular cytotoxicity assay.

J Immunol Methods 2019 May 18;468:49-54. Epub 2019 Feb 18.

Department of BioAnalytical Sciences, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080-4990, USA. Electronic address:

Antibody-dependent cellular cytotoxicity (ADCC) is an important mechanism of action (MOA) of monoclonal antibody (mAb) therapeutics. Target cells opsonized with therapeutic antibody bind and activate FcγR-bearing immune effector cells, resulting in target cell lysis. A key step in mAb drug development is the characterization of ADCC activity for its potential to inform mAb efficacy and safety. Read More

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http://dx.doi.org/10.1016/j.jim.2019.02.006DOI Listing

A dual neutrophil-T cell purification procedure and methodological considerations in studying the effects of estrogen on human Th17 cell differentiation.

J Immunol Methods 2019 Apr 13;467:1-11. Epub 2019 Feb 13.

Program in Translational Medicine, Peter Gilgan Centre for Research and Learning, The Hospital for Sick Children Research Institute, Canada; Departments of Paediatrics and Physiology, University of Toronto, Canada; Institute of Medical Sciences, University of Toronto, Canada. Electronic address:

New procedures are required to optimize the use of blood samples to study different cell types. The purification of neutrophils and T cells from the same blood sample is not commonly described. We have previously used PolymorphPrep™ (P) or LymphoPrep™ (L) for purifying neutrophils or T cells, respectively. Read More

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http://dx.doi.org/10.1016/j.jim.2019.01.002DOI Listing
April 2019
1 Read

The enzymatic removal of immunoglobulin variable domain glycans by different glycosidases.

J Immunol Methods 2019 Apr 8;467:58-62. Epub 2019 Feb 8.

Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands; Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands.

About 15% of immunoglobulin G (IgG) molecules contain glycans linked to the antigen-binding fragments (Fab arms) in addition to the glycans linked to the crystallizable fragment (Fc tail) of all IgGs. Fab glycosylation appears to be an important feature of antibodies, for example by influencing antigen binding and antibody stability. The reliable generation of antibodies that either have or lack Fab glycans would be very helpful to study the role of Fab glycans in more detail. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183043
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http://dx.doi.org/10.1016/j.jim.2019.02.005DOI Listing
April 2019
1 Read

A monoclonal antibody targeted to the functional peptide of αB-crystallin inhibits the chaperone and anti-apoptotic activities.

J Immunol Methods 2019 Apr 6;467:37-47. Epub 2019 Feb 6.

Sue Anschutz-Rodgers Eye Center and Department of Ophthalmology, University of Colorado School of Medicine, United States; Skaggs School of Pharmacy and Pharmaceutical Sciences, Anschutz Medical Campus, Aurora, CO 80045, United States. Electronic address:

αB-Crystallin is a member of the small heat shock protein family. It is a molecular chaperone and an anti-apoptotic protein. Previous studies have shown that the peptide (DRFSVNLDVKHFSPEELKVKV, hereafter referred to as peptain-1) from the core domain of αB-crystallin exhibits both chaperone and anti-apoptotic properties similar to the parent protein. Read More

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http://dx.doi.org/10.1016/j.jim.2019.02.004DOI Listing
April 2019
1 Read

An improved clonogenic culture method for thymic epithelial cells.

J Immunol Methods 2019 Apr 6;467:29-36. Epub 2019 Feb 6.

Department of Immunology and Cell Biology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan; Center for iPS Cell Research and Application (CiRA), Laboratory of Immunobiology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan. Electronic address:

A clonogenic assay system for thymic epithelial cells (TECs) is of crucial importance for identifying thymic epithelial stem and/or progenitor cells, evaluating their activities, and understanding the mechanisms of thymic involution. However, current systems are not sufficiently sensitive at detecting and quantifying TEC colonies from the adult thymus. Here, we optimized the culture condition to detect visible colonies from adult TECs by modifying our previous culture methods. Read More

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http://dx.doi.org/10.1016/j.jim.2019.02.003DOI Listing
April 2019
1 Read

A novel method to efficiently isolate medullary thymic epithelial cells from murine thymi based on UEA-1 MicroBeads.

J Immunol Methods 2019 Apr 5;467:12-18. Epub 2019 Feb 5.

AG Clinical Immunology, Institute of Immunology, Rostock University Medical Center, Rostock, Germany.

Objective: The central mechanism for establishing a self-tolerant and functional T cell repertoire includes the promiscuous expression of otherwise tissue-restricted proteins by medullary thymic epithelial cells (TEC). We here demonstrate a novel and highly efficient method for isolating this rare key cell type.

Methods: We combined the enrichment of medullary TEC via UEA-1 MicroBeads with the subsequent depletion of residual CD45 hematopoietic cells via specific size exclusion and compared our results to the standard Percoll enrichment method and isolation procedure via flow cytometric cell sorting. Read More

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http://dx.doi.org/10.1016/j.jim.2019.02.001DOI Listing

Optimized protocols for studying the NLRP3 inflammasome and assessment of potential targets of CP-453,773 in undifferentiated THP1 cells.

J Immunol Methods 2019 Apr 5;467:19-28. Epub 2019 Feb 5.

Inflammation & Immunology Research Unit, Pfizer, Cambridge, MA 02139, United States. Electronic address:

The NLRP3 inflammasome is a complex multimeric signaling apparatus that regulates production of the pro-inflammatory cytokine IL-1β. To overcome both the variability among primary immune cells and the limitations of genetic manipulation of differentiated human or murine macrophages, we developed a simplified, reliable and relevant cell-based model for studying the NLRP3 inflammasome using the undifferentiated human myelomonocytic cell line THP1. Undifferentiated THP1 cells constitutively express NLRP3, and NLRP3 inflammasome activation occurred in response to canonical NLRP3 activation stimuli including nigericin, ATP, and urea crystals, culminating in pro-IL-1β cleavage, extracellular release of mature IL-1β, and pyroptosis. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183040
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http://dx.doi.org/10.1016/j.jim.2019.02.002DOI Listing
April 2019
21 Reads

Optimisation, harmonisation and standardisation of the direct mycobacterial growth inhibition assay using cryopreserved human peripheral blood mononuclear cells.

J Immunol Methods 2019 Jan 31. Epub 2019 Jan 31.

The Jenner Institute, University of Oxford, UK.

A major challenge to tuberculosis (TB) vaccine development is the lack of a validated immune correlate of protection. Mycobacterial growth inhibition assays (MGIAs) represent an unbiased measure of the ability to control mycobacterial growth in vitro. A successful MGIA could be applied to preclinical and clinical post-vaccination samples to aid in the selection of novel vaccine candidates at an early stage and provide a relevant measure of immunogenicity and protection. Read More

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http://dx.doi.org/10.1016/j.jim.2019.01.006DOI Listing
January 2019
1 Read

Development and validation of HIV-1 Multiplex Serology.

J Immunol Methods 2019 Mar 18;466:47-51. Epub 2019 Jan 18.

Infections and Cancer Epidemiology, Infection, Inflammation and Cancer Research Program, German Cancer Research Center (DKFZ), Heidelberg, Germany; Faculty of Biosciences, Heidelberg University, Heidelberg, Germany. Electronic address:

By inducing immunosuppression in infected patients, human immunodeficiency virus-1 (HIV-1) generates a favorable environment for opportunistic infections and the development of several human cancers. In order to detect individual serum or plasma HIV-1 antibody status for epidemiological studies, high-throughput HIV-1 Multiplex Serology was developed. Seven HIV-1 antigens were recombinantly expressed in E. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183035
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http://dx.doi.org/10.1016/j.jim.2019.01.007DOI Listing
March 2019
6 Reads

A novel tandem repeat cloning technique for creation of multiple short peptide repeats to differentiate closely related antigens.

J Immunol Methods 2019 Jan 17. Epub 2019 Jan 17.

Microbiology Division, Defence Food Research Laboratory, Siddarthanagar, Mysore, Karnataka 570011, India.

Antibody cross-reactivity is a problem often associated with closely related antigens. This study was aimed to develop a method enabling differentiation of closely related toxins based on antigen designing strategy. The method involves identification of disparate amino acids (AA) confined to target antigen in comparison with two or more closely related antigens, their assembly into a DNA oligomer and further cloning as six tandem repeats (TR) using restriction and ligation strategy into a desired vector and finally generation of antigen specific antibodies. Read More

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http://dx.doi.org/10.1016/j.jim.2019.01.008DOI Listing
January 2019
2 Reads

Development and validation of different indirect ELISAs for MERS-CoV serological testing.

J Immunol Methods 2019 Mar 16;466:41-46. Epub 2019 Jan 16.

Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia; Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.

Since 2012, MERS-CoV has caused up to 2220 cases and 790 deaths in 27 countries with Saudi Arabia being the most affected country with ~83.1% of the cases and ~38.8% local death rate. Read More

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http://dx.doi.org/10.1016/j.jim.2019.01.005DOI Listing
March 2019
5 Reads
2.005 Impact Factor

Reducing the risk of misdiagnosis of indirect ELISA by normalizing serum-specific background noise: The example of detecting anti-FGFR3 autoantibodies.

J Immunol Methods 2019 Mar 14;466:52-56. Epub 2019 Jan 14.

Synaptopathies and Autoantibodies, Faculty of Medicine Jacques Lisfranc, University Jean Monnet, University of Lyon, Saint-Étienne, France; Synaptopathies and Autoantibodies, Institut NeuroMyoGene INSERM U1217/CNRS UMR 5310, University of Lyon, Université Claude Bernard Lyon 1, Lyon, France; Neurology Department, Centre Hospitalier Universitaire de Saint-Étienne, Saint-Étienne, France.

Indirect enzyme-linked immunosorbent assay (ELISA) is an important diagnostic method as it enables the quantification of the presence of autoantibodies in human blood sera. However, unspecific binding of antibodies to the solid phase causes considerable serum-specific background noise (SSBN), involving the risk of false positive diagnosis. Therefore, we present a simple and concise, yet obvious proof-of-principle of a recently suggested normalization method. Read More

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http://dx.doi.org/10.1016/j.jim.2019.01.004DOI Listing
March 2019
2 Reads

Determination of human γδ T cell-mediated cytotoxicity using a non-radioactive assay system.

J Immunol Methods 2019 Mar 14;466:32-40. Epub 2019 Jan 14.

Center for Bioinformatics and Molecular Medicine, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Program for Nurturing Global Leaders in Tropical and Emerging Infectious Diseases, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Center for Innovation in Immunoregulative Technology and Therapeutics, Graduate School of Medicine, Kyoto University, Yoshidakonoe-Cho, Sakyo-Ku, Kyoto 606-8501, Japan; Hyogo College of Medicine, 1-1 Mukogawa, Nishinomiya, Hyogo 663-8501, Japan. Electronic address:

The adoptive transfer of immune effector cells, such as CD8 killer αβ T cells, γδ T cells, NK (natural killer) cells, and genetically-modified T cells, has been receiving increasing attention. It is essential to determine cellular cytotoxicity so as to monitor the function and quality of ex vivo-expanded immune effector cells before infusion. The most common method is the [Cr]-sodium chromate release assay. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183039
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http://dx.doi.org/10.1016/j.jim.2019.01.003DOI Listing
March 2019
5 Reads

Development of a modified yeast display system for screening antigen-specific variable lymphocyte receptor B in hagfish (Eptatretus burgeri).

J Immunol Methods 2019 Mar 3;466:24-31. Epub 2019 Jan 3.

Laboratory of Aquatic Animal Diseases, Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju, Gyeongnam 660-701, South Korea. Electronic address:

The variable lymphocyte receptor B (VLRB) of jawless vertebrates has a similar function to the antibodies produced by jawed vertebrates, and has been considered as an alternative source to mammalian antibodies for use in biological research. We developed a modified yeast display vector system (pYD8) to display recombinant hagfish VLRB proteins on the extracellular surface of yeast for the isolation of antigen-specific VLRBs. After observing an up-regulation in the VLRB response in hagfish immunized with hemagglutinin 1 of avian influenza virus H9N2 subtype (H9N2-HA1), the antigen-specific VLRBs decorated on the yeast's surface were selected by quantitative library screening through magnetic-activated cell sorting (MACS) and fluorescent-activated cell sorting (FACS). Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183039
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http://dx.doi.org/10.1016/j.jim.2019.01.001DOI Listing
March 2019
15 Reads

Measurement of IL-21 in human serum and plasma using ultrasensitive MSD S-PLEX® and Quanterix SiMoA methodologies.

J Immunol Methods 2019 Mar 24;466:9-16. Epub 2018 Dec 24.

Immunology Discovery, Eli Lilly and Company, Indianapolis, IN 46285, USA.

IL-21 is a pleiotropic cytokine that plays a key role in modulating inflammatory responses, including the promotion of autoimmune diseases. Several groups have quantitated circulating levels of IL-21 in plasma and serum samples using various commercial ELISAs. We determined, however, that the most commonly used commercial assays in published literature were not specific or sensitive enough to detect levels of IL-21 in heparin plasma or serum from healthy human individuals. Read More

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http://dx.doi.org/10.1016/j.jim.2018.12.005DOI Listing
March 2019
8 Reads

Optimization of single-cell plate sorting for high throughput sequencing applications.

J Immunol Methods 2019 Mar 24;466:17-23. Epub 2018 Dec 24.

Stanford University, Department of Medicine/Nephrology, Palo Alto, CA 94304, United States; VA Palo Alto Health Care System, Palo Alto, CA 94304, United States. Electronic address:

Single cell sequencing has recently been applied to many immunological studies. Flow cytometric index sorting isolates cells for single cell sequencing with protein level data linked to sequences. However, successful sequencing of index sorted samples requires careful optimization of several sort parameters, including nozzle size, flow rate, threshold rate, and yield calculations. Read More

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http://dx.doi.org/10.1016/j.jim.2018.12.006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6363834PMC
March 2019
3 Reads

Phage-based assay for rapid detection of bacterial pathogens in blood by Raman spectroscopy.

J Immunol Methods 2019 Feb 12;465:45-52. Epub 2018 Dec 12.

Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Viale F. Stagno d'Alcontres 31, 98166 Messina, Italy. Electronic address:

Sepsis is a systemic inflammatory response ensuing from presence and persistence of microorganisms in the bloodstream. The possibility to identify them at low concentrations may improve the problem of human health and therapeutic outcomes. So, sensitive and rapid diagnostic systems are essential to evaluate bacterial infections during the time, also reducing the cost. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183038
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http://dx.doi.org/10.1016/j.jim.2018.12.004DOI Listing
February 2019
16 Reads

An optimized protocol for the analysis of house dust mite (Dermatophagoides farinae)-induced neutrophil-dominant airway inflammation.

J Immunol Methods 2019 Feb 12;465:53-60. Epub 2018 Dec 12.

Department of Immunology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan; AMED-CREST, AMED, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan. Electronic address:

House dust mites (HDMs), Dermatophagoides sp., are one of the most widespread aeroallergens worldwide and cause various allergic diseases, including asthma. The pathophysiology of asthma has been intensively investigated using murine models of allergic airway inflammation induced by exposure to D. Read More

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http://dx.doi.org/10.1016/j.jim.2018.12.003DOI Listing
February 2019
6 Reads

The relevance of buffer system ionic strength in immunoassay development.

Authors:
Axel Petzold

J Immunol Methods 2019 Feb 11;465:27-30. Epub 2018 Dec 11.

Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London WC1N 3BG, United Kingdom. Electronic address:

The best validated immunoassays for neurodegeneration have been developed for class III and IV intermediate filaments. There are a number of unique biochemical features of the intrinsically unstructured polyampholytic tail regions of these proteins which affect domain structure and thereby affinity and epitope recognition of antibodies used in immunoassays. Here one of these intermediate filaments, the neurofilament heavy chain, is chosen to demonstrate the effect of the ionic strength of a buffer system on the analytical signal to noise ratio. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183035
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http://dx.doi.org/10.1016/j.jim.2018.11.013DOI Listing
February 2019
19 Reads

Simple, rapid and inexpensive typing of common HLA class I alleles for immunological studies.

J Immunol Methods 2019 Feb 8;465:72-76. Epub 2018 Dec 8.

University of Queensland Diamantina Institute, The University of Queensland, Translational Research Institute, Woolloongabba, Queensland, Australia. Electronic address:

Current HLA-typing methods are typically designed to provide exquisitely-detailed identification of multiple HLA-alleles to satisfy the requirements for organ and bone marrow transplantation or genetic studies. Many human immunological studies, on the other hand, focus around only a small number of HLA alleles that are abundant or of relevance to specific diseases. Consequently, for such studies, many HLA typing approaches are not cost-effective and are potentially complicated, slow and not easily performed in-house. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183020
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http://dx.doi.org/10.1016/j.jim.2018.12.002DOI Listing
February 2019
12 Reads

High-sensitivity lateral flow immunoassay with a fluorescent lanthanide nanoparticle label.

J Immunol Methods 2019 Feb 4;465:39-44. Epub 2018 Dec 4.

Department of Biotechnology, University of Turku, Turku, Finland.

Lateral flow (LF) immunoassays are commonly used for point-of-care testing and typically incorporate visually read reporters, such as gold particles. To improve sensitivity and develop quantitative LF immunoassays, visual reporters can be replaced by fluorescent reporters detected by an instrument. In this study, we used fluorescent europium(III) chelate doped nanoparticle (Eu-np) reporters to develop a quantitative high-sensitivity LF immunoassay for free prostate specific antigen (fPSA). Read More

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http://dx.doi.org/10.1016/j.jim.2018.12.001DOI Listing
February 2019
2 Reads

Production of a recombinant monoclonal antibody to Herpes Simplex Virus glycoprotein D for immunoaffinity purification of tagged proteins.

J Immunol Methods 2019 Feb 28;465:31-38. Epub 2018 Nov 28.

Department of Biomolecular Engineering, The University of California at Santa Cruz, Santa Cruz, CA, USA. Electronic address:

We have developed a stable Chinese Hamster Ovary (CHO) cell line for the production of a recombinant monoclonal antibody (mAb) to a short protein sequence derived from the N-terminus of human herpes simplex virus type 1 glycoprotein D (HSV-1 gD). The antibody (designated r34.1) provides a useful tool for the immunoaffinity purification of HSV-1 gD tagged proteins, and provides a generic purification system by which various proteins and peptides can be purified. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.015DOI Listing
February 2019
16 Reads

Cross-species reactive monoclonal antibodies against the extracellular domains of the insulin receptor and IGF1 receptor.

J Immunol Methods 2019 Feb 27;465:20-26. Epub 2018 Nov 27.

Global Drug Discovery, Novo Nordisk, Måløv, Denmark. Electronic address:

Translation across species of immunoassay results is often challenging due to the lack of cross-species reactivity of antibodies. In order to investigate the biology of insulin and IGF1 receptors, we generated new versatile monoclonal assay antibodies using the extracellular domain of the insulin/IGF1 hybrid receptor as the bait protein in the Adimab yeast antibody discovery platform and as the antigen in a rabbit monoclonal antibody platform. The resulting antibody clones were screened for receptor specificity as well as cross-species reactivity to both tissue and cell line derived samples. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183028
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http://dx.doi.org/10.1016/j.jim.2018.11.014DOI Listing
February 2019
6 Reads

IL8 and EDEM3 gene expression ratio indicates peripheral blood mononuclear cell (PBMC) quality.

J Immunol Methods 2019 Feb 26;465:13-19. Epub 2018 Nov 26.

Integrated Biobank of Luxembourg, 1 rue Louis Rech, 3555 Dudelange, Luxembourg; International Society for Biological and Environmental Repositories (ISBER), Biospecimen Science Working Group, Canada. Electronic address:

Background: Uncontrolled preanalytical variables can reduce the accuracy and reproducibility of downstream analytical results from peripheral blood mononuclear cells (PBMCs).

Methods: PBMCs were isolated from EDTA and citrate-anticoagulated blood samples, obtained from healthy subjects and patients with inflammatory and infectious conditions. PBMC-derived RNA samples were examined for gene expression changes induced by extended blood pre-centrifugation delays at 4 °C and RT. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.012DOI Listing
February 2019
4 Reads

A primer set for comprehensive amplification of V-genes from rhesus macaque origin based on repertoire sequencing.

J Immunol Methods 2019 Feb 22;465:67-71. Epub 2018 Nov 22.

Bioengineering Faculty of Engineering, Bar-Ilan University, Ramat Gan, Israel.

Recombinant antibodies serve as therapeutic molecules for a broad range of applications. High affinity antibodies are typically isolated following an active and effective immunization. Human-like antibodies may be obtained from immunized nonhuman primates (NHP), such as rhesus macaque, when immunized human origin is not available. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.011DOI Listing
February 2019
3 Reads

Improving TCR affinity on 293T cells.

J Immunol Methods 2019 Mar 22;466:1-8. Epub 2018 Nov 22.

Division of Immune Response, Aichi, Cancer Center Research Institute, Nagoya 464-8681, Japan; Division of Cellular Oncology, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan. Electronic address:

This study presents an efficient method to improve TCR affinity, comprising 1) CDR-directed saturation mutation of TCR cDNA, 2) transient TCR display on CD3-expressing HEK293T (CD3-293T) cells by simple plasmid transfection, 3) staining with HLA-tetramers, and 4) multi-round sorting of cells with CD8-independent tetramer binding on a flow cytometer. Using these procedures, we successfully identified mutant TCRs with enhanced binding from an HLA-A*24:02-restricted, human telomerase reverse transcriptase (hTERT)-specific TCR. Two such clones, 2A7A and 2D162, harboring mutations in CDR1 and CDR2 of TCRβ, respectively, were isolated with both showing sequential four amino acid substitutions. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.010DOI Listing
March 2019
19 Reads

Unravelling enhancement of antibody fragment stability - Role of format structure and cysteine modification.

J Immunol Methods 2019 Jan 22;464:57-63. Epub 2018 Nov 22.

School of Biotechnology, Dublin City University, Dublin 9 D09 Y5N0, Ireland,; Qatar Foundation, Hamad Bin Khalifa University, Education City, Doha, Qatar.

Antibody-based diagnostics and therapeutics have huge commercial value. However, applications of antibodies are often limited by instability, particularly for recombinant antibody formats. This paper describes the conversion of a single-chain variable fragment (scFv) antibody to a single-chain antibody fragment (scAb) with notably improved stability characteristics. Read More

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http://dx.doi.org/10.1016/j.jim.2018.10.012DOI Listing
January 2019
4 Reads

Non-invasive dynamic monitoring initiation and growth of pancreatic tumor in the LSL-Kras;LSL-Trp53;Pdx-1-Cre (KPC) transgenic mouse model.

J Immunol Methods 2019 Feb 20;465:1-6. Epub 2018 Nov 20.

Department of Radiology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA; Robert H. Lurie Comprehensive Cancer Center, Chicago, IL 60611, USA. Electronic address:

The LSL-Kras;LSL-Trp53;Pdx-1-Cre (KPC) mouse is one of the most widely used transgenic models to evaluate tumor characteristics and to develop novel therapies for pancreatic ductal adenocarcinoma (PDAC). There is no report of the effective systemic evaluation of longitudinal KPC tumor imitation and growth. Therefore, we aimed to characterize the initiation and progression of pancreatic cancer in KPC mice using longitudinal multiparametric magnetic resonance imaging (MRI) approaches and overall survival. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183038
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http://dx.doi.org/10.1016/j.jim.2018.11.009DOI Listing
February 2019
12 Reads

Comparison of bead array and glass nanoreactor multi-analyte platforms for the evaluation of CNS and peripheral inflammatory markers during HIV infection.

J Immunol Methods 2019 Feb 20;465:7-12. Epub 2018 Nov 20.

Department of Medicine, University of California at San Diego School of Medicine, La Jolla, CA, United States; Department of Psychiatry, University of California at San Diego School of Medicine, La Jolla, CA, United States.

While human immunodeficiency virus (HIV) infection has become a treatable disease with the development of combination antiretroviral therapy (cART), chronic inflammation that affects the central nervous system and other organs is still common. Reliable methods are needed to study HIV-associated inflammatory biomarkers. In this study involving both plasma and cerebrospinal fluid (CSF), we compared multiplex bead array (MBA) to a relatively new technology based on microfluidics and glass nanoreactor (GNR) technology for the measurement of three commonly studied markers from HIV-infected individuals. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.008DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6456896PMC
February 2019
15 Reads

Updates in diagnostic and clinical laboratory immunology from the 30th annual meeting of the Association of Medical Laboratory Immunologists (AMLI).

J Immunol Methods 2019 Jan 16;464:138-140. Epub 2018 Nov 16.

Pediatrics and Pathology, Keck School of Medicine, University of Southern California, Laboratory Medicine, Children's Hospital of Los Angeles, CA, USA.

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http://dx.doi.org/10.1016/j.jim.2018.11.007DOI Listing
January 2019
3 Reads

Effect of peripheral blood mononuclear cell cryopreservation on innate and adaptive immune responses.

J Immunol Methods 2019 Feb 14;465:61-66. Epub 2018 Nov 14.

Pneumococcal Research, Murdoch Children's Research Institute, Melbourne, Melbourne, VIC 3052, Australia; Department of Paediatrics, University of Melbourne, Australia. Electronic address:

Cryopreservation of blood-derived immune cells is commonly used in clinical trials to examine immunological responses. However, studies elucidating the effects of cryopreservation on peripheral blood mononuclear cell (PBMC) responses have shown inconsistent results making it difficult to draw meaningful conclusions. Therefore we sought to address this issue by comparing key innate and adaptive immune parameters between freshly-isolated and cryopreserved PBMCs from healthy adults. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183030
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http://dx.doi.org/10.1016/j.jim.2018.11.006DOI Listing
February 2019
16 Reads